Structure and Function of Paxillin

项目摘要 这项研究计划的广泛持续目标是更深入地了解 粘着斑适配器/支架蛋白桩蛋白和Hic-5 有助于细胞基质信号传导，从而控制细胞极性和发育中的迁移， 疾病重要的是，细胞骨架的三个要素之间的沟通，运输 机械和细胞-基质相互作用对于建立顶-基和前-后 迁移极性然而，我们对协调和整合的关键机制的理解 这些进程仍然没有完成。通过我们开发新的桩蛋白和Hic-5敲除 小鼠模型，结合使用各种离体1D、2D和3D细胞基质和类器官 模型系统和真实的时间成像方法，我们已经确定了桩蛋白的新作用， 建立上皮和间充质细胞极性，包括调节中心体和高尔基体 通过控制HDAC6活性来控制微管的组织和稳定性。我们还发现Hic-5是一种 一种新的F-actin介体-中间丝串扰、机械生物学和三维细胞外基质 沉积和重塑。将这些模型系统与定量的真实的时间结合使用 共焦，光片和超分辨率成像方法，我们将在 目标1：桩蛋白如何有助于调节极化 定向间充质细胞迁移的交易目标2：桩蛋白在建立 在乳腺上皮细胞的分支形态发生和目标3： HIC-5对运动细胞中波形蛋白中间丝细胞骨架结构和功能的调节 在上皮-间质转化过程中呢通过这些机制的阐明，我们将 更有能力制定合理的疾病干预方法， 发育障碍

PROJECT SUMMARY The broad ongoing objective of this research program is to gain a deeper understanding of the fundamental mechanisms by which the focal adhesion adaptor/scaffold proteins paxillin and Hic-5 contribute to cell-matrix signaling and thereby control of cell polarity and migration in development and disease. Importantly, communication between the three elements of the cytoskeleton, the trafficking machinery and cell-matrix interactions is essential for establishing both apical-basal and front-rear migration polarity. However, our understanding of the key mechanisms coordinating and integrating these processes remains incomplete. Through our development of new paxillin and Hic-5 knock out mouse models, in combination with the use of various ex-vivo 1D, 2D and 3D cell matrix and organoid model systems and real time imaging approaches, we have identified new roles for paxillin in establishing epithelial and mesenchymal cell polarity, including regulation of centrosome and Golgi organization and microtubule stability via control of HDAC6 activity. We have also identified Hic-5 as a new mediator of F-actin-intermediate filament cross talk, mechanobiology and 3D extracellular matrix deposition and remodeling. Using these model systems in conjunction with quantitative real time confocal, light sheet and super resolution imaging approaches, the main goals that we will address in the upcoming funding period are- Goal 1: How does paxillin contribute to the regulation of polarized trafficking in directed mesenchymal cell migration? Goal 2: What is the role of paxillin in establishment of apical-basal polarity and in branching morphogenesis in mammary epithelial cells and Goal 3: How does Hic-5 regulate the vimentin intermediate filament cytoskeleton organization and function in motile cells and during epithelial-mesenchymal transition? Through the elucidation of these mechanisms we will be better positioned to develop rational approaches to disease intervention and to appreciate the basis of developmental disorders.